A peptide carrier for the delivery of biologically active proteins into mammalian cells
A peptide carrier for the delivery of biologically active proteins into mammalian cells
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DOI：
10.1038/nbt1201-1173
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发表时间：
2001-12-01
影响因子：
46.9
通讯作者：
Divita, G
中科院分区：
文献类型：
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作者：
Morris, MC;Depollier, J;Divita, G
细胞膜的渗透性差和选择性差限制了肽类药物和治疗性蛋白质的开发。通过设计将全长蛋白质递送到大量细胞中的策略来规避这些问题的努力越来越多（1-3）。一系列小的蛋白质结构域，称为蛋白质转导结构域（PTD），已经显示出有效地穿过生物膜并且独立于转运蛋白或特异性受体，并且促进肽和蛋白质递送到细胞中。来自人免疫缺陷病毒（HIV-1）的达特蛋白能够在体内递送生物活性蛋白，并且已经显示对于蛋白质治疗具有相当大的意义（4-9）。类似地，双足同源结构域的第三个α-螺旋（10-12）和来自单纯疱疹病毒的VP 22蛋白（13，14）促进共价连接的肽或蛋白质递送到细胞中.然而，这些PTD载体显示出一定数量的限制，因为它们都需要与靶肽或蛋白质交联。此外，使用PTD-TAT融合蛋白系统的蛋白转导可能需要在递送前使蛋白变性以增加TAT-PTD结构域的可接近性。这一要求在递送时间和蛋白质的细胞内活化之间引入了额外的延迟（1）。在这份报告中，我们提出了一个新的策略，蛋白质输送的基础上短的两亲性肽载体，Pep-1。这种肽载体能够以完全生物活性的形式有效地将各种肽和蛋白质递送到几种细胞系中，而不需要预先的化学共价偶联或变性步骤。此外，这种肽载体呈现出蛋白质治疗的几个优点，包括在生理缓冲液中的稳定性、缺乏毒性和缺乏对血清的敏感性。Pep-1技术对于靶向活细胞中的特异性蛋白质-蛋白质相互作用和筛选新型治疗蛋白质非常有用。
The development of peptide drugs and therapeutic proteins is limited by the poor permeability and the selectivity of the cell membrane. There is a growing effort to circumvent these problems by designing strategies to deliver full-length proteins into a large number of cells(1-3). A series of small protein domains, termed protein transduction domains (PTDs), have been shown to cross biological membranes efficiently and independently of transporters or specific receptors, and to promote the delivery of peptides and proteins into cells. TAT protein from human immunodeficiency virus (HIV-1) is able to deliver biologically active proteins in vivo and has been shown to be of considerable interest for protein therapeutics(4-9). Similarly, the third alpha -helix of Antennapedia homeodomain(10-12), and VP22 protein from herpes simplex virus(13,14) promote the delivery of covalently linked peptides or proteins into cells. However, these PTD vectors display a certain number of limitations in that they all require crosslinking to the target peptide or protein. Moreover, protein transduction using PTD-TAT fusion protein systems may require denaturation of the protein before delivery to increase the accessibility of the TAT-PTD domain. This requirement introduces an additional delay between the time of delivery and intracellular activation of the protein(1). In this report, we propose a new strategy for protein delivery based on a short amphipathic peptide carrier, Pep-1. This peptide carrier is able to efficiently deliver a variety of peptides and proteins into several cell lines in a fully biologically active form, without the need for prior chemical covalent coupling or denaturation steps. In addition, this peptide carrier presents several advantages or protein therapy, including stability in physiological buffer, lack of toxicity, and lack of sensitivity to serum. Pep-1 technology should be extremely useful for targeting specific protein-protein interactions in living cells and for screening novel therapeutic proteins.